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Kansas State University

Flow Cytometry Lab


The Flow Cytometry laboratory is housed in Mosier Hall, Room K208 of the College of Veterinary Medicine Complex. This multi-user core laboratory is equipped with state of the art flow cytometry analyzers by Sony (EC800) and Becton Dickinson (FACSCalibur), a multiplex MagPix instrument, and fluorescent activated cell sorting equipment by Beckman Coulter (MoFLO XDP).

Hours of Operation: M – F 8 – 5 pm, Call to schedule a time on the instruments

Usage Fees:

INSTRUMENT

Price/ 15min

Price/ 30min

BD FACs CALIBUR

$20

 

Sony EC800

$25

 

Assisted flow on Analyzers

$30

 

BioRad S3 Cell Sorter

 

$45*

Beckman MoFlo XDP Cell Sorter

 

$50*

MagPix Setup (by staff)

$30/ setup

 

TRAINING PACKAGES (2 hrs)

$100

 

* Sorters will be charged a minimum of 30 minutes, and are operated by core staff only

New users will be strongly encouraged to sign up for a minimum of 2 hours of training.  Training will cost $100, and will accommodate up to 2 trainees per session.  The training sessions will cover instrument set-up, data acquisition, trouble-shooting, and introduction to analysis software. 


E-mail or call for consultation:

Director:

Melinda J. Wilkerson, DVM PhD

wilkersn@vet.k-state.edu

Ph: 785-532-4818

Research Assistant Professor and Associate Director:

Catherine Ewen, PhD

 cewen@vet.k-state.edu 

Ph: 785-532-4617
FAX Number: 785-532-4072

Research Assistant (Flow Cytometry certified operator):

Anushka George, MS

ageorge@vet.k-state.edu

Ph: 785-532-4617
FAX Number: 785-532-4072

 

INSTRUMENT

LASERS

FLUORESCENT MARKERS

 

FACS CALIBUR

 

488nm, 642nm

FITC, GFP, Alexa 488, PE, PI, PerCP, 7-AAD, Pe-Cy7, APC,   Alexa 647

 

BioRad S3 Sorter

 

488nm, 561 nm

FITC, GFP, Alexa 488, PE, PI, PerCP, 7-AAD, RFP, DsRed, mCherry

 

Sony EC800

 

405nm, 488nm, 561nm, 642nm

Brillaint Violet spectrum, DAPI, Pacific Blue, FITC, GFP, Alexa 488, PE, PI, PerCP, 7-AAD, RFP, DsRed, mCherry, APC, Alexa 647, Pe-Cy7, Alexa 700, PerCP Cy5.5

 

MoFlo XDP Sorter

 

355nm, 405nm, 488nm, 561nm, 642nm

Hoechst Blue, Brillaint Violet spectrum, DAPI, Pacific Blue, FITC, GFP, Alexa 488, PE, PI, 7-AAD, RFP, DsRed, mCherry, APC, Alexa 647, Pe-Cy7, Alexa 700, PerCP Cy5.5

 

Sony EC800:

 


The Sony EC800 is equipped with four lasers (405, 488, 561, and 640) with 6-color capabilities. This instrument can determine size, complexity, and electronic volume.  This flow cytometer can sample directly from a variety of plate or tube formats (ependorf tubes, falcon tubes, culture well plates, and tube rack) and is fully automated. The 405 laser allows the flexibility to use violet dyes. The 561 lasers provide the capability to excite a variety of fluorescent proteins (mCherry, mKate, link to fluorescent pdf chart).

FACSCalibur:



The FACSCalibur is equipped with two lasers that emit light at 488 nm and 630 nm. There are three detectors (FL1, FL2, FL3) for the 488 laser and one detector (FL4) for the 630 laser.  This two laser cytometer allows the operator to analyze up to 4-color and 2 size parameters of cells or fluorescent-labeled particles. FACSCalibur is interfaced with a Macintosh computer and a publication quality color printer. Cell Quest and ModFit analysis software is available on two computers to analyze flow cytometry data including DNA and proliferation assays.

BioRad S3:

BioRad S3

The BioRad S3 is equipped with two lasers (488 and 561) with 4-color capabilities. The 561 lasers provide the capability to excite a variety of fluorescent proteins. This instrument can determine size, complexity, and sort cells.  The sorting operation is fully automated.  This instrument will be placed in the BRI in a level 2-biosafety hood (BioBubble).

MoFLO XDP:

MoFLO XDP

The MoFLO XDP is a “state of the art” analytical flow cytometer with high speed sorting capabilities. The MoFlo has five lasers (UV/violet/argon/green/red) with the capability to perform 10-color and 2 size parameter analyses and to routinely sort cells or particles at speeds up to 70,000 cells/second.

The research capabilities of this facility are very diverse and only limited by the imagination, available fluorochromes, and capabilities of the instruments.  Examples of the research experiments that can be performed at this facility include but are not limited to: apoptosis studies, immunophenotyping, DNA content and viability of spermatozoa and other cell types, intracellular calcium levels, respiratory burst and phagocytic function, cell cycle, enzymatic activity, sorting of individual subpopulations of cells from a mixed population, sorting of GFP expressing cells from mixed cultures, and analysis of PCR products attached to microspheres. (Link to ppt slides for basic theory and applications)

MagPix:



The MagPix by Millipore can analyze up to 50 analytes in a single sample.  Examples are cytokine, chemokine, and serological panels. Call for consultation and quote for usage.

http://www.millipore.com/bmia/flx4/bmia_instruments#tab1=2

Additional educational resource (Tutorials on flow cytometry)

http://probes.invitrogen.com/resources/education/tutorials/4Intro_Flow/player.html